Background: Small GTPases are a super-family of cellular signaling regulators. Rac belongs to the Rho sub-family of GTPases that regulate cell motility, cell division, and gene transcription. GTP binding increases the activity of Rac, and the hydrolysis of GTP to GDP renders it inactive. GTP hydrolysis is aided by GTPase activating proteins (GAPs), while exchange of GDP for GTP is facilitated by guanine nucleotide exchange factors (GEFs).
Immunogen: Recombinant full length protein of active Rac1
Applications: IP, IHC
Recommended Dilutions: 1 µg for 1~2 mg total cellular proteins for IP;
1:100 dilution for IHC
Concentration: 1 mg/ml
Host: mouse
Format: Liquid
Clonality: Monoclonal
Isotype: IgM
Purity: Purified from ascites
Storage buffer:
Preservative: no
Constituents: PBS (without Mg2+ and Ca2+), pH 7.4, 150 mM NaCl, 50% glycerol
Species Reactivity: Anti-active Rac1 antibody recognizes active Rac1 from vertebrates.
Storage Conditions: Store at -20°C. Avoid freeze / thaw cycles
Immunoprecipitation/Western blot:
Rac activation assay. MEF cells were treated with (lane 2) or without (lane 1) PDGF. Cell lysates were incubated with an anti-active Rac monoclonal antibody (Cat # 26903) (top panel). Th precipitated active Rac was immunoblotted with an anti-Rac rabbit polyclonal antibody (Cat # 21003). The bottom panel shows the Western blot with anti-Rac of the cell lysates used (5% of that used in the top panel).
Immunofluorescence:
Immunofluorescence for the active Rac using Anti-Active Rac1-GTP Mouse Monoclonal Antibody [26903] shows Rac-GTP immunolabeling (green) in combination with cofilin (red) on brain tissue sections.
The tissue sections were fixed with -20 ºC methanol or 4% paraformaldehyde (fixation time 1hr) and stained with antibody at 1:1000 in 0.1M Phosphate buffer with 0.3% Triton X, and 4% BSA for 24h at room temperature. Secondary antibodies were anti-mouse AlexaFluor488 and anti-rabbit AlexaFluor594 at 1:1000.
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